• 대한바이러스학회지
• /
• 제26권1호
• /
• pp.59-65
• /
• 1996

For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

• KSBB Journal
• /
• 제13권6호
• /
• pp.656-661
• /
• 1998

Hantaan virus belonging to the genus Hantavirus and family Bunyaviridae causes an acute severe illness of human, Haemorrhagic Fever with Renal Syndrome (HFRS). It is a rodent host-borne pathogen and distributed in Asia and Eastern Europe. Hantaviruses have three major antigens, i.e., G1, G2 glycoproteins and nucleocapsid protein (N). Among them, nucleocapsid protein was reported to be the most invaluable antigen as for diagnosis. We have cloned and expressed Hantaan viral nucleocapsid gene in E. coli BL21(DE3). In this study, we have tried to purify the nucleocapsid protein produced by recombinant E. coli, and could attained a purity of >90% by anti-N monoclonal antibody-coupled immunoaffinity chromatography or phenyl sepharose hydrophobic interaction chromatography.

• Journal of Microbiology and Biotechnology
• /
• 제12권6호
• /
• pp.882-889
• /
• 2002

Hantaan vims production, using human immortalized retina cell (PER. C6), was investigated to develop an inactivated virus vaccine. To infect Hantaan virus into PER. C6, two infection methods (medium-to-cell and cell-to-cell) were tried, and IFA results showed that the cell-to-cell infection method was very useful for producing Hantaan virus-infected PER, C6. Hantaan virus production was significantly affected by the growth rate of PER. C6 and the content of FBS in medium. Higher specific growth rate of infected PER. C6 and lower FBS content induced higher production of Hantaan virus. The inactivated human cell-culture vaccines with various EIA titers were prepared, their antibody responses were compared with those of inactivated suckling mouse brain vaccines ($Hantavax^처리불가$). and the result showed their immunogenicities were slightly higher than those of inactivated suckling mouse vaccines. Therefore, this study shows the possibility of the development of Hantaan virus vaccine from a human cell culture.

• 대한바이러스학회지
• /
• 제28권1호
• /
• pp.11-20
• /
• 1998

Twenty isolates of Hantavirus were isolated from patients and reserovirs from 1988 to 1994 in Korea. Isolation rate was 1.9% (10/538) in patients, 6.2% (5/81) in Apodemus sp., 2.6% (1/38) in Rattus sp. and 0.6% (4/677) in bats. Reciprocal mean IFA titers ranged from 27.5 to 1,024 at the specimen collection. According to the growth rate and reaching peak titier of infectivity, the isolates were grouped as rapid, intermediate, and slow growing groups. All isolates were confirmed as Hantaan type by the nested RT-PCR on the G1 region of the M segment. Comparison of nucleotide sequence (Nt: 2101 - Nt: 2280) of the G2 region revealed that the sequence homology bewteen Hantaan 76/118 virus and the isolates was more than 90%. Several nucleotide positions of the isolates showed high variation. The variation rate of patientisolates was about one-half when compared with that of rodentisolates. On the basis of phylogenetic analysis Hantaan viruses isolated were divided into two genogroups. These results indicate that Hantaan virus is highly dominant serotype in Korea and the virologic property and genogroup are not correlated.

• Genomics & Informatics
• /
• 제6권1호
• /
• pp.54-58
• /
• 2008

Genotyping Tool for Viral SEQuences (GTVseq) provides scientists with the genotype information on the viral genome sequences including HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2, poliovirus, enterovirus, flavivirus, Hantavirus, and rotavirus. GTVseq produces alternative and additive genotype information for the query viral sequences based on two different, but related, scoring methods. The genotype information produced is reported in a graphical manner for the reference genotype matches and each graphical output is linked to the detailed sequence alignments between the query and the matched reference sequences. GTVseq also reports the potential 'repeats' and/or 'recombination' sequence region in a separated window. GTVseq does not replace completely other well-known genotyping tools such as NCBI's virus sequence genotyping tool (http://www.ncbi. nlm.nih.gov/projects/genotyping/formpage.cgi), but provides additional information useful in the confirmation or for further investigation of the genotype(s) for the newly isolated viral sequences.

• 대한바이러스학회지
• /
• 제27권1호
• /
• pp.59-68
• /
• 1997

Hantaan virus Howang strain which isolated from the blood of severe case of Korean hemorrhagic fever is more virulent than HTN 76/118 and showed different RFLP from partial PCR amplifed M genome segment to established Hantaan serotype viruses. We have determined the nucleotide sequence of the M and S genome segments and compared to HTN 76/118. The M and S segment of Howang strain has 3615 and 1696 nucleotides long, respectively. The M segment sequence of Howang strain is one mucleotide shorter than HTN 76/118. The sequence data of Howang strain shows 93.5% homology to HTN 76/118. One long open reading frame, which strats from 41nt. to 3448nt. of the M segment and from 37nt. to 1326nt. of the S segment, exist to on complementary sense of the virus genome. There are no significant difference between HTN 76/118 and Howang strain on hydrophobicity of deduced polypeptides, but has slight difference on secondary structure.

• 대한바이러스학회지
• /
• 제28권4호
• /
• pp.327-335
• /
• 1998

Hantaan (HTN) and Seoul (SEO) viruses, murid rodent-borne hantaviruses, are known to causes hemorrhagic fever with renal syndrome (HFRS) in Korea. To determine the genomic diversity and molecular phylogeny of HTN and SEO viruses found in Korea, we amplified for part of M and S genomic segments of hantaviruses from sera of HFRS patients and lung tissues of hantavirus seropositive striped-field mice. Both M and S segment of 16 HTN and 2 SEO viruses were amplified by nested reverse transcription-polymerase chain reaction. Based on 324 nucleotides in the M genomic segment, the HTN and SEO strains showed $93.8{\sim}100%$ and $99.1{\sim}99.4%$ homologies, respectively. Similarly, based on 230 nucleotides in the S genomic segment, HTN and SEO strains showed $90.9{\sim}100%$ and 100% homologies, respectively. Phylogenetic analysis of M and S segments indicated that HTN strains could be divided into at least two main groups in M and S trees and the sequence differences detected among the Sand M genomic segments of HTN viruses are consistent with reassortment having taken place between HTN virus strains.

• 대한의생명과학회지
• /
• 제16권4호
• /
• pp.311-316
• /
• 2010

We studied a comparison of the concentration of biochemical markers in sera of patients hospitalized with high fever (n=296) in Jeonbuk province during the last 2 years (2008 to 2009). The patients were divided into three patient groups of viral hemorrhagic fever (VHF) patient group tested positive for Hantavirus (n=53), leptospirosis (LEP) patient group tested positive for Leptospira interrogans (n=137) and scrub typhus (TSU) patient group tested positive for Orientia tsutsugamushi (n=106). We analyzed the concentration of ALP, AST, ALT, blood urea nitrogen, creatinine and glucose and compared the mean levels of them to normal range, the first sample and last sample. The frequencies of abnormal patient elevated above the upper limit of normal for ALP, AST and ALT were 18~43.4%, 78~97% and 62.3~92.7% in patient groups, and 24.5~47.4% (total protein) and 13.2~50.0% (albumin) of patients in patient groups had decreased below the lower limit of normal. The patients showed higher abnormal levels of glucose in patient groups were 58.5% (viral hemorrhagic fever patient group), 66.4% (leptospirosis patient group), 71.7% (scrub typhus patient group) and 66.9% (total patient group). There were significant difference between the first sample and the last sample in the mean levels of AST (decreased 22.2% in viral hemorrhagic fever patient group, 30.2% in leptospirosis patient group, 20.4% in scrub typhus patient group and 24.1% in total patient group), BUN (43.0% in viral hemorrhagic fever patient group, 41.6% in leptospirosis patient group, 47.4% in scrub typhus patient group and 43.0% in total patient group) and glucose (20.2% viral hemorrhagic fever patient group, 17.9% in leptospirosis patient group, 18.6% in scrub typhus patient group and 18.9% in total patient group) in the first sample and the last sample. According to these results, those diseases may cause liver damage and have high concentration of ALP, AST, ALT and glucose in blood even though the patients get out of the hospital.

• 대한바이러스학회지
• /
• 제29권4호
• /
• pp.201-209
• /
• 1999

Results of the studies on the morphologic and molecular biologic characteristics of Hantaan virus (HTNV), one of the etiologic agents of Hemorrhagic fever with renal syndrome (HFRS), revealed that HTNV was a member of Family Bunyaviridae and its RNA divided into three segments. And the nucleotide sequences of these segments also were known and the differences in nucleotide sequences of HTNV from other members of genus Hantavirus were clearly evaluated. But the morphorgenesis, pathogenesis of HFRS and the replication time had not been clearly determined. In this study, to estimate the replication time of HTNV in Vero E-6 cells, Vero cells were infected with HTNV 76/118 strain, and cells were harvested from two hours post-infection up to 24 hours at two hours-intervals. Harvested cells were treated with ordinary techniques for electron microscopy and immune-electron microscopy. And then thin sections were observed under transmission electron microscope. HTNV particles were not found in the cytoplasm and in the extracellular space between $2{\sim}8$ hours after inoculation of virus, but virus particles were observed in extracellular space near the cell membrane of Vero-E6 cells 10 hours after infection. In immune electron microscopy, mature HTNV particles in extracellular spaces and immature virus labelled with gold particles in the cytoplasm of Vero E-6 cell 10 hours after infection of HTNV could be seen. This results suggest that the replication time of HTNV might be about 10 hours.

• 대한바이러스학회지
• /
• 제29권4호
• /
• pp.221-233
• /
• 1999

Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.